FI60555C - AERATING CARBOXYLAMIDE DERIVATIVES FOR FRAMSTATING OF THERAPEUTIC ANALYSIS - Google Patents

Description

ΓβΙ ms ADVERTISEMENT 6 055 5 jSjjj IBJ <11> UTLÄGGNINGSSKRIFT CUO ° 0 C Patent granted tty 10 C £ 1 '0 ^ Patent oeddelat ^ ^ (51) Kv.ik.3 / int.ci.3 C 07 C 103/84 FINLAND — FINLAND (21) Pttcnttlhaktmui - Pit «nMns6knlnf 1 ^ 5/7 ^ (22) Hakamltptlvt - AntAknlngsdig I8.OI.7I + ^ (23) Alkupltvt — GHti (het * d» | l8.Ol.7lt

(41) Interpreters | ulktMkil - Bllvlt off «ntllg 19.07.7U

National Board of Patents and Registration .... ωΜ..Μ .., _ *. (44) Date of NihtlvIk »lp» noo and moonshine. -

Patent and ocean registration in the field of trade and services »krtft * n pubixerad 30.10.8l (32) (33) (31) Pyydetty« uelkau * —Begirt priority 18.01.73

Japan-Japan (JP) 7359 / Sho h8 (71) Kissei Yakuhin Kogyo Kabushiki Kaisha, No. 105 Nomizo Yoshikawa-ku, Matsumoto-shi, Nagano-ken, Japan-Japan (JP) (72) Kozaburo Harita, Nagano-ken, Yukiyoshi Ajisawa, Nagano-ken, Kinji Iizuka, Nagano-ken, Yukihiko Kinoshita, Nagano-ken , Tetsuhide Kami jo, Nagano-ken, Michihiro Kobayashi, Nagano-ken, Japan-Japan JP) (7M Oy Kolster Ab (5lt) Method for the preparation of therapeutically useful aromatic carboxylamide derivatives - Förfarande för framställning av tera-peutisktest More particularly, this invention relates to a process for the preparation of substituted cinnamoylaminobenzoic acid derivatives and substituted hydrocinnamoylaminobenzoic acid derivatives having a strong antiallergic effect when administered orally to mammals in humans.

Until now, disodium cromoglycate has been the only drug that inhibits mast cell rupture and the release of chemical mediator compounds in it. However, this compound loses its pharmacological effect when ingested orally and the range to which this compound can be applied is naturally limited. Thus, the development of an antiallergic agent that can demonstrate adequate therapeutic efficacy when taken orally has long been demanded in the medical field.

2 60555

Reinicke, on the other hand, had already synthesized cinnamoylaminobenzoic acid, the phenyl moiety of which is unsubstituted and is well known (Liebing's Annaler der Chemie, Vol. 3 ^ + 1, pages 9 ^ -96) · However, this compound proves to have only a very weak anti-allergic effect when used is administered orally to mammals and therefore was hardly useful as a practical medicine.

Accordingly, it is an object of the present invention to provide a compound having a potent anti-allergic effect when administered orally to mammals, including man.

Another object of the present invention is to provide a novel aromatic carboxylamide derivative having a pharmacological effect.

Another object of the present invention is to provide cinnamoylaminobenzoic acid and hydrocinnamoylaminobenzoic acid having substituted phenyl moieties, and physiologically acceptable salts thereof.

Other objects, features and advantages of the present invention will become apparent from the following. The attached figure shows the change in physiological state over time when a compound of this invention is administered to rats experimentally infected with asthma.

It has now been found that a compound of the general formula fy-? - r s - (I) JRiR2 0 M!

(x) COOH

n wherein and represents a hydrogen atom or an alkyl group having 1 to 1 carbon atoms, R 1 and represent a hydrogen atom or together form a chemical bond, the substituents X are the same or different and represent a hydroxyl group or an alkoxy group having 1 to 1 carbon atoms, and n is an integer 2 or 3, can inhibit experimentally induced sensitization (e.g., inflammation of the skin caused by an antigen-antibody reaction between a reagent and a specific antigen) when administered orally to patients. It has also been found that this compound inhibits mast cell rupture caused by a particular antigen-antibody reaction (e.g., an antigen-antibody reaction between a reacting antibody and a specific antigen) and the subsequent release of chemical mediators from the mast cell.

3 60555 In light of these characteristics, it is expected that this compound will have an anti-allergic effect and will be effective in the therapeutic treatment of diseases caused by allergies such as asthma, hay fever, articaria and natural dermatitis.

In fact, this compound was found to be effective in relieving the symptoms of respiration and blood pressure observed in experimentally induced asthma in mammals.

The compounds of this invention are characterized in that they have at least two substituents on the phenyl of the cinnamoyl group, which may be hydroxy or alkoxy groups. If the compounds do not have such substituents, they become weak in their anti-allergic effect and less valuable for practical purposes. In case these substituents are alkoxy groups they can be straight or branched. To the extent that the number of carbon atoms in such groups is 1-¼, no significant change in pharmacological potency is observed. The number of substituents is limited to 2-3.

In the compounds of this invention, the position of the carboxyl group in the aminobenzoic acid residue may be any of the 2-, 3- and k-positions. Salts of compounds containing this carboxyl group, such as alkali metal salts, are as potent in pharmacological activity as the corresponding compounds having free acid groups, whereas compounds in the form of their esters with lower alcohols have been found to be inferior in pharmacological activity.

According to the invention, the compounds of formula (I) are prepared by a process in which a reactive derivative of an aromatic carboxylic acid of formula II, R 3 R 1 * C 1 -C 3 C-COOH (II) (X) n 11 60555 wherein R 1, R 2, R 2 »X and n have the same meaning as above, are reacted with an aminobenzoic acid of the formula: —COOH (III)

V

nh2 or by a process in which an aromatic carboxylic acid of the above general formula (II) or a reactive derivative thereof is reacted with an ester of an aminobenzoic acid having the general formula:? - COOR.

V

nh2 wherein Rg is an alkyl group to give an aromatic carboxylamide derivative of the general formula RR, ^ '3'k -cc-NH-rn (im 11 "v 1 1 R, R0 0 1 2 (Xin C00E6 where R1, R2> R 3, R 2, R 9, X and n have the same meaning as above, and then this derivative is hydrolyzed to convert the ester group to a carboxyl group, or a malonylic acid derivative of the formula N 2 - nhcoch 2 cooh (IV)

COOH

60555 is condensed with an aromatic aldehyde of formula

acho

wherein X and n are as defined above, to give a compound of formula I wherein ^ forms a chemical bond, and if desired, the resulting amide is converted to the corresponding salt.

The aromatic carboxylic acids of the above general formula (II) are known compounds and can be easily prepared according to methods described in the literature. Aromatic carboxylic acids with an unsaturated bond comprise two isomers, i. the cis form and the trans form, both of which can be used in the process of this invention. Examples of aromatic carboxylic acids of general formula (II) are 2,3-dimethoxycinnamic acid, 3'-dimethoxy-cinnamic acid, 4-methyl-3,3-dimethoxycinnamic acid, β-methyl-3'-dimethoxycinnamic acid, 1-diethoxycinnamic acid and 2,1,5-trimethoxycinnamic acid. In the process of the present invention, reactive derivatives of such aromatic carboxylic acids are used as a starting material. Examples of such derivatives include carboxylic acid derivatives such as acid halides, acid anhydrides, mixtures of acid anhydrides and esters, and reactions of said carboxylic acid and carbodiimide. These reactive derivatives can be readily derived from the aromatic carboxylic acids of general formula (II) according to a conventional method known in the art. For example, acid chlorides can be easily obtained by refluxing an aromatic carboxylic acid with thionyl chloride for several hours without any solvent or in dry benzene. Esters can be obtained by heating a particular aromatic carboxylic acid with an alcohol in the presence of an acid. A mixture of acid halides can be obtained, for example, by reaction with a chloroformic acid ester.

Any anthranilic acid, m-aminobenzoic acid and p-aminobenzoic acid can be used as the aminobenzoic acid of general formula (III). Any anthranilic acid ester, m-aminobenzoic acid ester and p-aminobenzoic acid ester can be used as the aminobenzoic acid ester of general formula (IIIJ).

The above-mentioned amidation can be carried out according to a method known in the art. For example, when an acid halide is used as a reactive derivative, the acid halide can be reacted in an inert solvent with an aminobenzoic acid of general formula (lii) or a derivative of general formula (lii ') in the presence of a basic substance, in this case a tertiary organic base or , such as alkali hydroxide, sodium carbonate or potassium carbonate are used as the basic substance. Suitable inert solvents include chloroform, methylene chloride, acetone, benzene, toluene, tetrahydrofuran, dioxane and dimethylformamide.

Instead of using such a basic substance, the reaction may be carried out using an excess of a compound of general formula (III), e.g. more than twice the molar amount of a compound of general formula (II). The process of this invention is preferably carried out by dissolving a compound of general formula (III) in dry chloroform and dry pyridine. in which these solvents are present in 5 and 10 times and 2 to 15 times the amount of the compound of general formula (II), respectively, a solution of the compound of general formula (II) in dry chloroform is added to this mixture under cooling and stirring and then refluxing for several hours. .

The reaction product is concentrated under reduced pressure and the residue is poured into water. Chlorohydric acid is then added to the aqueous mixture to make it weakly acidic. The precipitated crystals are collected by filtration and then recrystallized from a sufficient amount of organic solvent to obtain the final product.

When the compound of the general formula (II) is an aromatic carboxylic acid having a hydroxyl group in the benzene nucleus, such a compound is preferably protected from its hydroxyl group by an acetyl group or the like before reacting with the compound of the general formula (II). Such a protecting group can be removed in the usual manner.

The obtained compound (i) containing a carboxyl group can be converted into a physiologically acceptable salt according to a conventional method. For example, an aqueous solution of sodium hydroxide may be added in an equimolar amount to an alcoholic solution of the compound of general formula (i) and the mixture heated to a time sufficient for the compound to be readily converted to its sodium salt. Examples of such physiologically acceptable salts include, in addition to said sodium salt, alkali metal salts such as potassium salt and lithium salt, alkaline earth metal salts such as magnesium salt and calcium salt, salts with organic amines such as piperidine, triethanolamine and diethylaminoethylamine.

In particular, the aromatic carboxylamide derivatives prepared according to the present invention have activity against the effect of the antigen-antibody reaction. Thus, they can be widely used as therapeutic drugs for allergies.

The present invention will be described in more detail by means of examples in which not all melting points of the products have been corrected.

Example 1

Anthranilic acid (1.8 g) was dissolved in a mixture of dry chloroform (25 ml) and dry pyridine (7 ml). To this mixture was added dropwise and under cooling 10 ml of dry chloroform containing 2.5 g of 2-acetoxy-3-methoxycinnamoyl chloride. The mixture was refluxed for 2 hours and the reaction mixture was concentrated under reduced pressure. The residue was poured into water, and the liquid was weakly acidified by adding chlorohydric acid. The precipitated crystals were collected by filtration and recrystallized from aqueous alcohol to give 2.1 g of 2- (2'-acetoxy-3'-methoxycinnamoylamino) benzoic acid, m.p. + 231-23l ° C.

Elemental analysis of the compound C ^ H O ^ N C H N

Calculated {%) 61+, 22 1+, 82 3.9 ^

Experimental {%) 61 +, 21 + 1 +, 87 3.99 IR absorption spectrum (KBr) Y CO: 1765, 1695, 1670 cm-1 NMR spectrum (d 6 -DMSO) δ: 2.1 + 1 (s, 3H , acetoxide) 3.81+ (Sj 3H, methoxyhydrogen) 6.90, 7.61+ (q5 2H, J = 16 Hz, olefinic hydrogen) 7.10-8.70 (m, 6h, aromatic ring hydrogen) 11, 1 + 2 (s, 1H, hydrogen carboxylic acid),

A mixture of 2.0 g of 2- (2'-acetoxy-3'-methoxycinnamoylamino) benzoic acid, 38 ml of 10% aqueous sodium hydroxide solution and 38 ml of ethanol was heated for 2.5 hours. The reaction mixture was concentrated under reduced pressure, cooled and made. weakly acidified with hydrochloric acid. The precipitated crystals were collected by filtration and recrystallized from aqueous alcohol to give 1.3 g of 2- (2J-hydroxy-3, -methoxycinnamoylamino) benzoic acid, m.p. 206-208 ° C.

8 60555

CHON Elemental Analysis 1 f 15 5

C H N

Calculated {%) 65.17 1+, 82 U

Experimental {%) 65.17 1+, 80 1+, 52 IR absorption spectrum (KBr) δCO: 1680, 1660 cm -1 NMR spectrum (d 6 -DMSO) δ · 3.87 (s, 3H, methoxide) 6.8 * +, 7.96 (q, 2H, J = 16 Hz, hydrogen olefin) 6.80-8.80 (m, 7H, hydrogen of the aromatic ring) 9.33 (broad, 1H, hydrogen amide) 11.38 (s, 1H, hydrogen carboxylic acid).

1.05 g of 2- (2'-hydroxy-3'-ethoxycinnamoylamino) benzoic acid was dissolved in 1,500 ml of heated ethanol. To this solution was added a solution of 0.1 L of sodium hydroxide in 1.7 ml of water, and the mixture was heated for 30 minutes. The reaction mixture was concentrated under reduced pressure, and the residue was recrystallized from aqueous alcohol to give 0.55 g of the sodium salt of 2- (2'-hydroxy-3'-methoxycinnamoylamino) benzoic acid, m.p. 2l + 8-25l + ° C (decomposes).

Elemental analysis of the compound C ^ H ^ O ^ NNa C H N

Calculated (%) 60.89 1+, 27 1+, 18

Experimental {%) 60.83 1+, 32 1+, 21 IR absorption spectrum (KBr) δ CO: 1655 cm -1 NMR spectrum (d 6 -DMSO) δ: 3.85 (s, 3H, methoxy hydrogen) 6.72, 7.78 (q, 2H, J = 16 Hz, olefinic hydrogen) 6.72-8.75 (m, 7H, aromatic ring hydrogen),

The following compounds can be prepared in a similar manner: 6 3 '{y' '.' PCsC-C-N-'N 5 »» “1 il)., 1., Ί H.R.0 H n: i.

? 1+

x »51 3 TOOH

Compound No. η X R ^ R ^ COOH - position of the group Sp. (° C) Crystallization solvent 1 2 2'-OH 3, -0Me HH 1+ 307-307.5 Alcohol water decomposes solution 2 2 l + '- OH 31 -OMe HH 2 230-232 Alcohol 3 2 l +' - OH 3 '-OMe HH 3 238-239,5 Aqueous solution of alcohol 1+ 2 3'-OH l +' - OH HH 2 20l + - 206 decomposes 5 2 l + '- OH 3'-OMe HH 1+ 250-251 9 60555

Example 2

In the same manner, except that 3-methoxy-U-acetoxyhydrocinnamoyl chloride was used instead of the phenyl-substituted cinnamoyl ii urissil used in Example 1 to react with 3-aminobenzoic acid and the reaction product was hydrolyzed, 3 - (3'-methoxy-N-hydroxyamylamine hydroxyl) was obtained. After recrystallization from aqueous alcohol, this product had a melting point · of 218-220 ° C,

Example 3 H, 3 g of U-aminobenzoic acid was dissolved in a mixture of 100 ml of dry chloroform and 19 g of dry pyridine. To this mixture was added dropwise and with cooling 100 ml of a dry chloroform solution containing 5 * * g of 3'-dimethoxy-cinnamoyl chloride. The mixture was refluxed for 1.5 hours and then the reaction mixture was concentrated under reduced pressure. The residue was poured into water, and hydrochloric acid was then added to make the liquid weakly acidic. The precipitated crystals were collected by filtration and recrystallized from alcohol to give>> 6 g of U- (3 ', U'-dimethoxycinnamoylamino) benzoic acid, m.p. 267-9 ° C.

Elemental analysis for C 18 H 28 N 2 O 2

C H N

Calculating (%) 66.05 5.2U U, 28

Experimentally (%) 66.00 5.12 U, 1U

IR absorption spectrum (KBr) KCO: 1690, 1665 cm -1 KNH: 3320 cm NMR spectrum (dg VdMSO) δ: 6.78, 7.62 (q, 2H, J = 16 Hz olefin hydrogen) 7.0-7 .3 (m, 3H, hydrogen of methoxy-substituted aromatic ring) 7.85, 7.97 (q, UH, J = 9 Hz, hydrogen of amino-substituted aromatic ring) 10, U (s, 1H, hydrogen of carboxylic acid) 11.5 -12.7 (broad, 1H, hydrogen amide) 3.81, 3.83 (ss, 6h, hydrogen methoxy)

Mass spectrum M +, 327 m / e: 282, 191, 163.

1.5 g of U- (3 *, U'-dimethoxycinnamoylaminoaminobenzoic acid) was dissolved in 150 ml of heated ethanol and an aqueous alcoholic solution (ethanol 2: water 1) containing an equimolar amount of sodium hydroxide was added, whereupon white crystals precipitated. dried under reduced pressure to give 1.0 g of sodium U- (3, -U'-dimethoxycinnamoylamino) benzoate.

ίο 6 0 5 5 5

In the same manner as described above, the following compounds can be prepared: 2 j 2 3 —c = c-c-n — yf \ k! '' "'\

h iVn / ^ τ> τ> r \ u ^ - C00H

h 6 <h1R2 ° H 6 5 X 5 'n

Compound No. X R 1 R 2 COOH group Sp. (° C) Crystallization solvent _location_ 1 2 21-OMe 3'-OMe HH 2 190.5-200 aqueous alcohol solution 2 2 2'-OMe 3'-OMe HH k 261-263.5 alcohol 3 2 3'-OMe k ' -OMe HH 2 211-213 chloroform 1+ 2 3'-OMe k'-OMe HH 3 225-226 alcohol 5 2 3'-OMe k'-OMe HH 1+ 267-269 "6 2 3'-OMe k ' -OMe CH 2 H 2 173-175 aqueous alcohol solution 7 2 3'-OMe k'-OMe CH 3 H 3 163 ~ 16k "8 2 3'-OMe k'-OMe CH3 H k 2kl-2k3 alcohol 9 2 3'- OMe h'-OMe H CH3 2 169-172 aqueous alcohol solution 10 2 3'-OMe k'-OMe H CH3 3 203.5 ~ 20k, 5 "11 2 3'-OMe k '-OMe H CH3 k 225.5 -227.5 methanol 12 2 3'-OEt k'-OEt HH 2 185-187 aqueous alcohol solution 13 2 3'-OEt k'-OEt HH k 253-256 "1k 2 k'-OEt 3'-OMe HH 2 210.5-212.5 15 2 k'-OEt 3'-OMe HH 3 219-222 "16 3 2'-OMe k -'- OMe HH 2 206-208" 5J-OMe 17 2 k'-On- Pr HH 2 172-177 chloroform 3'-OMe 18 2 k'-Oi-Pr HH 2 76-78 "3J-OMe 19 2 2'-OMe 3'-OMe HH 3 238-2k0 alcohol / ligroline 20 2 2 ' -0Me k'-OMe HH 2 188—191 aqueous alcohol solution 21 2 21-OMe 5 '“OMe HH 2 181-183” 60555

Example 4

In a similar manner, except that the cinnamoyl chloride used in Example 3 was replaced with the corresponding hydrocinnamoyl chloride, the following compounds could be prepared: 2 '2_3 3' ^ Yp ch-ch-c-n - \ 4

b2 h y = pC0 ° H

<< 5.

Compound No. X COOH group M.P. (° C) Crystallization solvent _location_ 22 2 31-OMe 4'-OMe H H 3 176-177 alcohol / benzene 23 2 3'-OMe 4'-OMe H H 2 136-137.5 benzene

Example 5 4 g of 3,4-dimethoxycinnamic acid were dissolved in 20 ml of dry pyridine.

To this solution was added 2 g of benzenesulfonyl chloride under ice-cooling and stirring to form a red-orange precipitate. The reaction mixture was stirred for about an hour, and then 2 g of methyl anthranilate was added to the mixture under ice-cooling. The mixture was then stirred for 2 hours at room temperature to complete the reaction. After completion of the reaction, the reaction mixture was concentrated, and the residue was dissolved in about 10 ml of chloroform. The solution was washed first with 10% aqueous sodium hydroxide solution, then with 10% aqueous hydrochloric acid solution, and finally with water, and then distilled to remove chloroform to give crystals of N- (3 ', 4J-dimethoxycinnamoyl) -anthranilic acid methyl ester.

This product was dissolved in 10 ml of ethanol. To this solution was added 10 ml of 10% aqueous sodium hydroxide solution, and the mixture was kept at 50 ° C to effect hydrolysis of the ester group. After completion of the reaction, the reaction mixture was acidified with dilute hydrochloric acid, and chloroform was added to the reaction mixture. The organic phase was separated, washed with water and distilled to remove the solvent to give 2.1 g (48% yield) of the final product, m.p. N- (3 *, 4'-dimethoxy-cinnamoyl) -antraniilihappoa. The melting point of this product was 211-213 ° C.

When the condensation reaction was carried out under similar conditions except that 1.9 g of anthranilic acid was used instead of the methyl anthranilate used in this example, 2.0 g of N- (31,4'-dimethoxycinnamoyl) -anitric acid could be obtained directly, 12 60555

Example 6 2 g of 3,4-dimethoxycinnamic acid were dissolved in a mixture of 20 ml of dry dimethylformamide and 1.5 g of triethylamine. To this solution was added 1.1 g of ethyl chlorocarbonate under ice-cooling and stirring, and the mixture was then allowed to react for one hour. To this mixture was added 10 ml of dimethylformamide containing 1.5 g of 3-aminobenzoic acid, and the mixture was stirred for 2 hours. After completion of the reaction, the reaction liquid was concentrated to approximately half its volume and poured into aqueous hydrochloric acid. The precipitated crystals were separated by filtration, washed with water and recrystallized from an equal volume of ethanol and water to give 3- (31,4'-dimethoxycinnamoylamino) benzoic acid with a melting point of 225-226 ° C in 50% yield. .

Example 7

To a mixture of 20 ml of dry dioxane and 0.7 g of dry pyridine were added 2 g of 3,4-dimethoxycinnamic acid, 1.6 g of phosphorus oxychloride and 1.6 g of methyl anthranilate. The mixture was refluxed for 2 hours to carry out the reaction. After completion of the reaction, the reaction mixture was concentrated, and the residue was dissolved by heating in ethanol and then cooled, whereupon crystals precipitated. The crystals were separated by filtration and treated in the same manner as in Example 5 to perform hydrolysis of the ester group. The product was recrystallized from chloroform to give 2 g of N- (31,4'-dimethoxycinnamoyl) -anthranilic acid, m.p. 211-213 ° C.

Example 8

A mixture of 3.2 g of 3,4-dimethoxybenzaldehyde and 4.6 g of malonanthranilic acid was added to 4 ml of pyridine and the mixture was heated at 80-100 ° C for three hours. The reaction mixture was evaporated in vacuo. The residue was dissolved in ethanol and the ethanol solution was poured into water. The aqueous mixture was acidified by the addition of hydrochloric acid. The precipitated crystals were collected and recrystallized from chloroform to give 5.3 g of 2- (3J, 4'-dimethoxycinnamoyl) benzoic acid, m.p. 212-216 ° C.

Elemental analysis of the compound C ^ H ^ O N

C H N

Calculated {%) 66.05 5.24 4.28

Experimental (%) 65.90 5.30 4.15 13 60555 IR absorption spectrum V CO: 1695, 1655 cm -1 NMR spectrum (d 6 -DMSO, 90 MHz) δ 3.02, 3.87 (s, s, 3H, methoxy hydrogen) 6.7-8.8 (m, 11H, hydrogen of aromatic ring, amide and olefin) 11.36 (s, 1H, hydrogen carboxylic acid)

Mass spectrum M + 327 m / e 309.191, 163.

Experiments "Equivalent passive skin sensitization in rats" Wistar male rats weighing 120-150 g were used in this experiment. The reactive antibody was obtained from rats immunized with egg white (EA) dissolved in pertussis-diphtheria-tetanus vaccine. Normal rats were passively sensitized by antibody dilution using intradermal injection. U8 hours after sensitization, a mixture of antigen different (EA) and Evans blue was injected intravenously. The animals were then sacrificed by hitting the head 30 minutes after injection and then the blue spot caused by the antigen-antibody reaction was measured photometrically.

The specified test compound dissolved in 1 # NaHCO 3 solution was orally administered at a dose of 200 mg / kg 2 hours before antigen injection, while the vehicle alone was administered to the control group. Chlorophenene used as a positive control is generally well known as an inhibitor of growth cell disruption caused by an allergic reaction.

The corresponding passive skin sensitization to control the efficacy of the test compounds was compared to a value (%) calculated using the following formula:

- x 100 A

where A is the amount of lost dye in the control group and B is the amount of lost dye in the group to which the particular test compound was administered.

It seems likely that a similar passive skin sensitization in rats is useful in determining whether or not a test compound suppresses an allergic reaction.

"· 60555

The results of the experiments were as shown below.

_Example No. _Compound_Excellent% _ 1 Comparison 0 2 Chlorophenene 36.7 3 N-Cinnamoylanthranilic acid 16.7 * + 3- (Cinnamoylamino) benzoic acid 10.0 5 * + - (Cinnamoylamino) benzoic acid 7.0 6 N - (* + '-hydroxycinnamoyl) anthranilic acid 36.7 7 3 ~ (3'-methoxycinnamoyl) benzoic acid * + 5.0 8 N- (2'-chlorocinnamoyl) anthranilic acid 60.2 9 N - (* +' - chlorocinnamoyl) anthranil acid 69, * + 10 * + - (* +, - bromocinnamoyl) benzoic acid 32.3 11 N- (3'-fluorocinnamoyl) anthranilic acid * + 7.5 12 N - (* + '-methylcinnamoyl) anthranilic acid 5 * +, 0 13 * + (2'-Hydroxy-3'-methoxycinnamoylamino) benzo [33.5] acid 1 * + N- (2'-3-dimethoxycinnamoyl) anthranilic acid 56.8 15 3- (2 ', 3'-dimethoxycinnamoylamino) ) benzoic acid * + 1,1 acid 16 N-3'-methoxy - * + '- n-propoxycinnamoyl) anthranilic 52.2 acid 17 N- (3'-methoxy - * +' - isopropoxycinnamoyl) anthranilic * + 7.1 acid 18 N- (3 ', * +' - dimethoxycinnamoyl) anthranilic acid * + 6.1 19 N- (2,3 * ', 5'-trimethoxycinnamoyl) anthranilic acid 56.8 20 N- (3 1, * + '- dimethoxyphenyl dihydroquinnamoyl) anthranilic acid 55, * + 21 N- (3 '- * +' -dimethoxy-N-methylcinnamoyl) anthra-66.2 nilic acid 22 K- (3'-methoxy - * + '- (2,3- dihydroxypropoxy) cinna- (9,0-moyl) -anthranilic acid 23 N- (3'-methoxy-* + - carboxylmethoxycinnamoyl) -0 anthranilic acid

The above results clearly show that the novel aromatic carboxylamide derivatives of this invention have almost the same or better pharmacological activity as chlorphenesin. Cinnamoylaminobenzoic acid derivatives substituted in the phenyl moiety are generally more potent than in the phenyl moiety. unsubstituted cinnamoylaminobenzoic acid derivatives. However, cinnamoylaminobenzoic acid derivatives substituted with a hydrophilic group in the phenyl moiety have virtually no activity.

15 60555 "Effect on sensitized mast cell disruption"

Intestinal mast cells isolated from normal rats were passively sensitized by germination with rat reactive antibody at 3 ° C. At the end of germination, specific antigen (DNP-Ascaris) was added to the germination medium, after which these brine feeder cells were solidified with formalin and stained with 0.1% toluidine blue (in acetic acid buffer; pH 1, 6). The number of mast cells disrupted by the antigen-antibody reaction was counted under a microscope. The number of disrupted mast cells was also counted without the addition of specific antigen.

A specific test compound dissolved in 1 # NaHCO 3 solution was added. . A concentration of 10 g / ml was added to the germination medium 5 minutes before antigen treatment, while only vehicle was added to the control group. Disodium bromoglycate, used as a positive control, is commonly known as an agent that inhibits mast cell rupture as a result of an allergic reaction.

The efficacy of test compounds in controlling mast cell rupture was compared to a value (%) calculated using the following formula: iPräHEiäi * 100 (P-Q) where P represents the percentage of ruptured mast cells in the control group, Q the percentage of ruptured mast cells, and R the percentage of ruptured mast cells.

It seems likely that this method is useful in determining whether or not a particular test compound inhibits mast cell rupture and subsequent release of chemical mediators from mast cells.

The test results were similar to those listed below.

Experiment No._Compound _Content% 1 Comparison 0 2 Disodium cromoglycate 16 3 N- (3''-dimethoxycinnamoyl) anthranilic acid 33 k N- (3J, -dimethoxy- / 1-methylcinnamoyl) anthranilic acid 5 5 3 ~ (h '-methoxycinnamoylamino) benzoic acid 18 6 N- (3'-methoxy-U'-n-propoxycinnamoyl) anthranilic acid 6 60555

The above results clearly show that the novel aromatic carboxylamide derivatives of the present invention have almost the same or better pharmacological activity as disodium cromoglycate.

"Experimentally induced asthma in rats" 20 normal male rats weighing 120-150 g were divided into four groups of 5 rats each. All rats were passively sensitized by intravenous injection of rat reactive antibody. Twenty-two hours after sensitization, rats in each group were orally administered 5 ml of an aqueous solution of 1-Ris NaHCO 3 containing 2- (3''-dimethoxycinnamoylamino) benzoic acid (Compound No. 33) at a dose of 0 mg / kg (comparison). ), 5 mg / kg, 10 mg / kg and 20 mg / kg, and then cannulation of both the trachea and the common main artery was started. Respiratory rate and volume as well as systemic blood pressure were determined simultaneously with a polygraph. After 2 hours of test substance administration, symptoms were induced by antigen injection and the results are shown in the drawing. Asthma symptoms occurred only in the control group. As shown in the drawing, respiratory rate decreased in the control group 3 minutes after antigen injection. On the other hand, Compound No. 33 had a inhibitory effect at a dose of 5 mg / kg and higher. With respect to the decrease in tidal volume, this compound had an inhibitory effect at a dose of 10 mg / kg and higher. The compound did not have an antihypertensive effect just after the antigen injection, but the compound prevented a subsequent reduction in blood pressure.

"Acute Toxicity" The mean lethal dose (LD50) of the novel aromatic carboxylic acid amide derivatives of this invention was determined by oral treatment in dd-male and female mice (6 weeks old) and in Wistar male and female rats (7 weeks old). and intraperitoneal treatment, Each animal selected for the experiment was kept in an air-conditioned room at a temperature of 22 + 1 ° C and a relative humidity of 55+ 5% for the duration of the experiment. All animals were fed compressed spheres (type CE-2, manufactured by Japan CLEA Co.) and water was available in the bottle as desired. Five groups of ten animals were used for each treatment.

Because these compounds are insoluble in water, they were suspended in a 0.5 # aqueous solution of carboxymethylcellulose in each aliquot. The treatment volume of each substance per 10 g of mouse body weight was 0.2 ml in the oral treatment, while it was 0.5 ml per 100 g of rat body weight in the intraperitoneal and oral treatment 'τ 60555 By the method of Litchfield-Wilcoxon. The results of the experiments were as follows: N- (3 ', 1 + J-dimethoxycinnamoyl) anthranilic acid Rats 1,850 mg / kg (p.o.) 2,030 "" 385 "(i.p.) 338" "

Mice 705 "(po) 500" N- (2'-chlorocinnamoyl) anthranilic acid Mice 3 ^ 2 "" N- (U'-chlorocinnamoyl) anthranilic acid Mice 551 "" N- (3'-chlorocinnamoyl) anthranilic acid Mice 700 "" N - (2'-fluorocinnamoyl) anthranilic acid Mice 321 "" N- (3'-fluorocinnamoyl) anthranilic acid Mice 551 "" N- (h'-fluorocinnamoyl) anthranilic acid Mice U81 ""

As shown in the experiments, the novel aromatic carboxylamide derivatives of this invention and their reactive derivatives work not only to prevent skin sensitization and the release of chemical mediators from mast cells, but also experimentally induced asthmatic symptoms caused by an antigen-antibody reaction.

This fact clearly supports the fact that these derivatives of this invention are effective in the therapeutic treatment of allergic diseases, including asthma, hay fever, articaria and natural dermatitis.

Claims (1)

18,60555 Claim; A process for the preparation of therapeutically useful aromatic carboxylamides and their pharmaceutically acceptable salts of the formula R, R 1, 3 C to C-CONH. -O X) ^ COOH where R. | and R 1 represents a hydrogen atom or an alkyl group having 1 to U carbon atoms, R 1 and R 2 represent a hydrogen atom or together form a chemical bond, the substituents X are the same or different and represent a hydroxyl group or an alkoxy group containing 1 to 1 carbon atoms, and n is an integer 2 or 3, characterized in that a) a mn-, R, R 5 reactive derivative of a capped aromatic carboxylic acid * 3, C - C-COOH II s of formula II, wherein R 1, R 2, R 2, X and n mean the same as above, is reacted with an aminobenzoic acid of formula i, _LL_cooh III V nh2 or b) an aromatic acid of formula II 19 60555? 3 C - C-COOH C '1 »II (X) - 1H, Rs" W A reactive derivative represented by the formula wherein R 1, R 2, R 2 and R 2, X and n are as defined above is reacted with an aminobenzoic acid ester of the formula with the formula - C-CONH ^^^. (X) n "f AKI -ft-C00R6 V wherein R1, R2, R1, R1, R8, X and n have the same meaning as above, after which the ester of formula 1 'is hydrolyzed to the free carboxylic acid compound of formula I, or c ) a malonylic acid derivative of the formula σΗ00 (ΙΗ2000Η IV COOH 20 6 0 5 5 5 is condensed with an aromatic aldehyde of the formula CHO ^ {v wherein X and n are as defined above to give a compound of formula I wherein and form a chemical bond, and, if desired, converting the amide obtained to the corresponding salt. In the case of a ring atom, R 1 and R 2 are a substituent or a substituent on a chemical bond, the substituent X is a residue or an oil or a hydroxyl group or an alkoxy group in the 1-U column. that heltal 2 or 3, the following is shown: a) that the reactive derivatives are derived from an aromatic carboxylic acid of formula II * 3 «u___ - C - COOH II with R ^, R2, R ^, R ^, X and n betecknar samma in the case of an amino-benzoic acid of formula - COOH III nh2 or b) that the reactants are derived from an aromatic syrup of formula II 22 60 5 5 5 R3 ru σο - C - COOH II '* for R. |, Rg, R ^ »R ^, X and n are selected from the group consisting of aramino-benzoic acid esters of formula I-jj- COORg III 'nh2 with Rg is an alkyl group, for which a carboxylamide derivative with the formula R- R.3 is used, UC - C - CONH »<X) n_R 'X ^^ __ C00Re r där R ^, Rg, R, R ^, Rg, X and n are selected from the group consisting of esters of formula 1' hydrolysed to the carboxylic acid formulation I, eller c) that malonanilidesyraderivat med med formeln Λ. NHCOCH COOH and COOH condensers with an aromatic aldehyde form